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Preparation of kit used for detecting benzodiazepine drug residues

A technology of benzodiazepines and kits, which is applied in the field of ELISA kits for detecting benzodiazepines, achieving the effect of sensitive detection and easy operation

Inactive Publication Date: 2016-02-10
JIANGSU WISE SCI & TECH DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, BZD determination methods mainly include high-performance liquid chromatography, gas chromatography, thin-layer chromatography, high-performance liquid chromatography-mass spectrometry, etc. The detection results are accurate and reliable, but the equipment is relatively expensive and the sample pretreatment is difficult. The process is relatively complicated

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  • Preparation of kit used for detecting benzodiazepine drug residues

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Embodiment Construction

[0023] Preparation of BZD protein conjugates:

[0024] Adopt succinic anhydride method to obtain the BZD hapten derivative with carboxyl group, then take 0.05mmol and carrier protein BSA according to the combination ratio of 10:1 and mix in the carbonate buffer solution (CBS) of 0.05mol / LpH9.6, then add 0.15mmol carbodiimide, stirred at room temperature for 24 hours, and finally dialyzed in 0.2mol / L PBS buffer solution of pH 7.6 for two days to remove unreacted haptens, and stored the obtained protein conjugate solution at -20°C spare.

[0025] Preparation of BZD antibody:

[0026] Select healthy adult purebred BALA / C mice, take 50 μg of immune antigen prepared by coupling with protein and mix with the same amount of complete Freund's adjuvant, and use intraperitoneal injection for initial immunization, and then use the same dose of immune antigen plus the same amount every 3 weeks Incomplete Freund's adjuvant was used for the second and third immunizations by intraperitonea...

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Abstract

The invention discloses an enzyme linked immunosorbent assay (ELISA) kit used for detecting benzodiazepine drug residues, and a detection method of the ELISA kit. The enzyme linked immunosorbent assay kit is high in detection sensitivity, accuracy, speed, and specificity; operation is simple; and the detection method is suitable for large-scale sample detection. The enzyme linked immunosorbent assay kit comprises an enzyme labeled plate coated with BZD antigens, a BZD standard substance, a BZD antibody working solution, a BZD enzyme labeled second antibody working solution, a substrate solution A, a substrate solution B, a stopping solution, a concentrated sample diluent, and a concentrated washing liquid. Solid phase indirect competitive enzyme-linked immune response is taken as the principle of the enzyme linked immunosorbent assay kit. According to the detection method, an extracted sample, the BZD enzyme labeled second antibody working solution, and the BZD antibody working solution are added into corresponding enzyme labeled holes for a certain time of incubation; after plate washing, the substrate solution A, and the substrate solution B are added, so that the color of an obtained mixture is changed to be blue under enzyme action; and the stopping solution is added, and the color is changed from blue to yellow. The shade of color developing and the content of BZD in the standard substance or the sample are inversely related. The direction method can be used for directly detecting the content of BZD in blood or tissue.

Description

technical field [0001] The invention relates to the technical field of ELISA detection, in particular to an ELISA kit for detecting benzodiazepines. Background technique [0002] BZD is a class of compounds with outstanding effects, which have clinical effects of anticonvulsant, anti-anxiety, muscle relaxation and hypnosis, and have less toxicity. In the 1970s, foreign countries did a lot of in-depth research on BZD appetite enhancers, tested the effects of 1500 compounds on animal feeding, and found that many BZD compounds can promote animal feeding, among which the effect of sulfazepam is the most significant. Intramuscular injection, gavage and dietary supplementation can effectively increase the food intake and frequency of healthy animals, especially ruminants, and increase the daily food intake. In recent years, some feed companies have randomly added sedative and hypnotic drugs to the feed in order to pursue the conversion rate of feed and high profits. The medical ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/531
Inventor 洪霞薛永来毛楠楠
Owner JIANGSU WISE SCI & TECH DEV